LOCUS Exported 7897 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Targeting expression to neuronal and glial cell types in mice, non-human primates, and human retina. ACCESSION . VERSION . KEYWORDS 217_pAAV-ProD25-CatCh-GFP-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7897) AUTHORS Juttner J, Szabo A, Gross-Scherf B, Morikawa RK, Rompani SB, Hantz P, Szikra T, Esposti F, Cowan CS, Bharioke A, Patino-Alvarez CP, Keles O, Kusnyerik A, Azoulay T, Hartl D, Krebs AR, Schubeler D, Hajdu RI, Lukats A, Nemeth J, Nagy ZZ, Wu KC, Wu RH, Xiang L, Fang XL, Jin ZB, Goldblum D, Hasler PW, Scholl HPN, Krol J, Roska B TITLE Targeting neuronal and glial cell types with synthetic promoter AAVs in mice, non-human primates and humans. JOURNAL Nat Neurosci. 2019 Jul 8. pii: 10.1038/s41593-019-0431-2. doi: 10.1038/s41593-019-0431-2. PUBMED 31285614 REFERENCE 2 (bases 1 to 7897) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7897 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 2211..2220 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2217..3140 /codon_start=1 /product="N-terminal fragment of channelrhodopsin-2 from the alga Chlamydomonas reinhardtii (Nagel et al., 2003)" /label=ChR2 /note="blue light-activated cation channel" /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN PSMLYLATGHRVQWLRYAEWLLTCPVICIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGA" CDS 3147..3863 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3192..3213) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3453..3472) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3800..3821 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3894..4482 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3947..3967) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4365..4376) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4514..4990 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4713..4732) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5030..5170 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5030..5159 /label=AAV2 ITR rep_origin 5245..5700 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5332..5351) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5542..5563 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5717..5736) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5836..5858 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5896..5914) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5982..6086 /gene="bla" /label=AmpR promoter CDS 6087..6947 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6305..6324) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7118..7706 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7607..7626 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 7768..7897 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 gcggccgcac gcgtattccg gtcacacggc caagattatt ccacctgcgc tttgagcaat 61 agggagaggg ctctggtgcc tcttcctgga atttgattaa ttcgcttgag tcagtcacag 121 aatttgagga agcattgata tttgaagatg tgttcttcta aaggatacaa atgaatatat 181 gcatagtgag agtttaggag atagagaggc aggccgagtt tgaggccagc ctggtctaca 241 caggcgttct aggagaacct gtctcatatg cacatgggcc tgggattaca catacaactg 301 caatggcagc acttggaaag cggaagcagg agaatcggaa tttcacactc atccttcatt 361 atagaagtcc aaacctgtgc taagctactt ggacatcgct aaaaaacagc aaaaatcttt 421 ctaggaattg ccaatgtata cacaccaagt tgtatttttg tagggcagac ttttccaact 481 tgccagatga taataatcat ttatattagc tacatttcag gctccagaat taacactgtt 541 actgaaatgt ataggttcta aaacatacca tttccaaata ttttaaagga ttaacatttt 601 tgaaaagcat gtgatcttcc aaccatattt tagggaacag acataagaag taggcaagtt 661 tgggaagaaa cagttcctga gggcatttct tcccaacctt gaatgccctg tgggttaact 721 gaggtcttgg taaaatgtag attatttact ggactttttg aactgagagt gcatttctaa 781 caaaactcca gggtatgtgg tccttgcatt acatatgggg tagcaacatt ctaaagcagt 841 gtttttcaac ctgtggatca agacccacag aagtcacata gcagatatcc tgcctattag 901 atatttacat taagattcag agcagtagca gaataggagt catcccaacc tgaggaactg 961 catcagagtc ccagcatcag gaagggtgag agccactgag ctaaagtcct ctaggtgagg 1021 gggctgcccc ggaaagactc atttaaatga aaccactgac acagagagct gacagatgag 1081 gtgggttccg tgtctgtgag gctctgctgg tcgtctcctc actcccatgg aagaaccacc 1141 gagatgaggg cgaggggcag agctaaccca gcctctaagt aggcagagtc tagtgtccag 1201 ctgcccaagg aagaagtttg ctgtgtgagg tggccctgat gtccgcacac acaaaatgcc 1261 agtgaagtct acttgaccaa gtgaagctgg tgtggaatgg gaagaagcac acacagtcag 1321 tctctctgca cacactctgt ccttcacttc ttcacttacc agagatttga tgagaaccta 1381 ctagcagatc agatttgatc cctgagtgga aaaaacgtac agtgggagat aaaagaggaa 1441 aacaacggat ctgagtttga ggttagccta gtctgagcaa gccggataca cagtaagacc 1501 ctgtctcaca tacatccact cgcacgcgcg cgcacacaaa cacacacaca cacacacaca 1561 cacacacaca cacacacaca catcgtgcta aggacaagat aggcatcctg agagatgagc 1621 caggacaaag aaccagtaat agctcctgga gcagcacatc tgttttgcca ggattatccc 1681 ttggatctct taaaaccgag accttgtaat ctgaagactc aacttgggct gtacccttaa 1741 ccttcagctc tatgatgcaa gtgagtccac aggaccggag gctttgagat gagcttttca 1801 gaagggagga gttggccgct tgctcccaga gctccagcac ctgcattctt ctggctatgt 1861 cagaagccag atcatttccc tcgttaaaaa caaaaacaaa aaaacaaaca aacaaaatgt 1921 tagtctttgc cctttatctg cctggcaaag cttttaattg gcttgatctg tcattccgct 1981 agacataaag gggacaatcc ccggattagg aaggagctct ccagctcggg taaggagtct 2041 caaggcaagg taggcaagca ccaccggtcc gcactctcgc ccagctttta cgggaagaag 2101 agaatgttac tctatcctaa catatttttc cttttcctct atctcacaga taggaaaaat 2161 ttaagagcca gagggaacgt cccttctcag aggagacagc agaaggatcc gccaccatgg 2221 attatggagg cgccctgagt gccgttgggc gcgagctgct atttgtaacg aacccagtag 2281 tcgtcaatgg ctctgtactt gtgcctgagg accagtgtta ctgcgcgggc tggattgagt 2341 cgcgtggcac aaacggtgcc caaacggcgt cgaacgtgct gcaatggctt gctgctggct 2401 tctccatcct actgcttatg ttttacgcct accaaacatg gaagtcaacc tgcggctggg 2461 aggagatcta tgtgtgcgct atcgagatgg tcaaggtgat tctcgagttc ttcttcgagt 2521 ttaagaaccc gtccatgctg tatctagcca caggccaccg cgtccagtgg ttgcgttacg 2581 ccgagtggct tctcacctgc ccggtcattt gcattcacct gtcaaacctg acgggcttgt 2641 ccaacgacta cagcaggcgc accatgggtc tgcttgtgtc tgatattggc acaattgtgt 2701 ggggcgccac ttccgccatg gccaccggat acgtcaaggt catcttcttc tgcctgggtc 2761 tgtgttatgg tgctaacacg ttctttcacg ctgccaaggc ctacatcgag ggttaccaca 2821 ccgtgccgaa gggccggtgt cgccaggtgg tgactggcat ggcttggctc ttcttcgtat 2881 catggggtat gttccccatc ctgttcatcc tcggccccga gggcttcggc gtcctgagcg 2941 tgtacggctc caccgtcggc cacaccatca ttgacctgat gtcgaagaac tgctggggtc 3001 tgctcggcca ctacctgcgc gtgctgatcc acgagcatat cctcatccac ggcgacattc 3061 gcaagaccac caaattgaac attggtggca ctgagattga ggtcgagacg ctggtggagg 3121 acgaggccga ggctggcgcg gtacccatgg tgagcaaggg cgaggagctg ttcaccgggg 3181 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 3241 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 3301 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 3361 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 3421 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 3481 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 3541 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 3601 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 3661 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 3721 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 3781 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 3841 tcggcatgga cgagctgtac aagtgataag aattcgatat caagcttatc gataatcaac 3901 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 3961 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 4021 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 4081 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 4141 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 4201 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 4261 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg 4321 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 4381 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 4441 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc 4501 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga 4561 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc 4621 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca 4681 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt 4741 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca 4801 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt 4861 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt 4921 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt 4981 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta 5041 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca 5101 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc 5161 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 5221 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg 5281 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg 5341 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 5401 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 5461 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 5521 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta 5581 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa 5641 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt 5701 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc 5761 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac 5821 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac 5881 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa 5941 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 6001 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 6061 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 6121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 6181 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 6241 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 6301 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 6361 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 6421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 6481 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 6541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 6601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 6661 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 6721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 6781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 6841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 6901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 6961 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 7021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 7081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 7141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 7201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 7261 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 7321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 7381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 7441 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 7501 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 7561 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 7621 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 7681 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 7741 ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg 7801 aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc 7861 agagagggag tggccaactc catcactagg ggttcct //